Study on the anti-inflammatory mechanism of moxibustion in rheumatoid arthritis in rats based on phospholipaseA2 signaling inhibition by Annexin 1.

Guo Yanding, Luo Kun, Zhang Linlin, L U Wenting, Shang Yanan, Zhong Yumei, H U Danhui, Yang Xin, Zhou Haiyan
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Abstract

Objective: To determine whether moxibustion had an anti-inflammatory effect on rheumatoid arthritis (RA) by regulating Annexin 1 expression and interfering with the phospholipaseA2 signaling pathway.

Methods: Thirty male Sprague-Dawley rats were randomly categorized into five groups (six rats per group): blank control (CON) group, RA model (RA) group, moxibustion (MOX) group, Annexin 1 lentiviral intervention (RNAi-Anxa1) group, and Annexin 1 lentiviral intervention + moxibustion (RNAi-Anxa1 + MOX) group. The rats in the RNAi-Anxa1 and the RNAi-Anxa1 + MOX groups were injected with the lentiviral vector-mediated RNAi-Anxa1 into the rat foot pad. An experimental RA rat model was established by injecting Freund's complete adjuvant (FCA) into the RA, MOX, RNAi-Anxa1, and RNAi-Anxa1 + MOX groups. Rats in the MOX and RNAi-Anxa1 + MOX groups received moxibustion treatment. After modeling, using moxibustion "Shenshu (BL23)" and "Zusanli (ST36)", each point is 5 times, bilateral alternating, once a day, 6 times for a course of treatment, between the courses of rest for a one day. A total of three treatment courses were conducted. Both bilateral pad thicknesses were measured using Vernier calipers on experimental days 1, 7, 14, 21, and 28. The expression of cPLA2α signaling in the synovium of diseased joints was observed using Western blot. The pathology of the rat ankle synovium was observed using hematoxylin-eosin (HE) staining. Interleukin (IL)-1β, IL-10, prostaglandin E2 (PGE2), and leukotriene B4 (LTB4) were detected using enzyme-linked immunosorbent assay.

Results: Moxibustion increased the levels of Annexin 1 and decreased the inflammatory response in rats with RA. After increasing the expression of Annexin 1, the phosphorylated expression of cPLA2α was inhibited, the serum levels of IL-1β, PGE2, and LTB4 decreased, and the level of IL-10 increased. In moxibustion treated RA rats after the Annexin 1 lentiviral intervention, the serum levels of IL-1β, PGE2, LTB4, and IL-10 were almost unchanged.

Conclusion: Moxibustion enhanced the negative regulation of the cPLA2α signaling pathway, increased the synovial Annexin 1 expression, inhibited the cPLA2α signaling pathway, indirectly inhibited the expression of downstream inflammatory factors, and played a role in reducing inflammation.

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基于Annexin 1抑制磷脂酶A2信号传导的艾灸对大鼠类风湿性关节炎抗炎机制的研究
目的方法:将30只雄性Sprague-Dawley大鼠随机分为5组(每组6只),分别为空白对照组(CON)、类风湿性关节炎模型组(RA)、艾灸组(RA):将30只雄性Sprague-Dawley大鼠随机分为5组(每组6只):空白对照(CON)组、RA模型(RA)组、艾灸(MOX)组、Annexin 1慢病毒干预(RNAi-Anxa1)组和Annexin 1慢病毒干预+艾灸(RNAi-Anxa1 + MOX)组。RNAi-Anxa1组和RNAi-Anxa1 + MOX组大鼠的足垫注射了慢病毒载体介导的RNAi-Anxa1。通过向RA组、MOX组、RNAi-Anxa1组和RNAi-Anxa1 + MOX组注射弗氏完全佐剂(FCA),建立了实验性RA大鼠模型。MOX 组和 RNAi-Anxa1 + MOX 组的大鼠接受艾灸治疗。建模后,用艾条灸 "神枢(BL23)"和 "足三里(ST36)",每穴均灸5次,双侧交替进行,每天1次,6次为1个疗程,疗程间休息1天。共进行三个疗程。在实验第 1、7、14、21 和 28 天,使用游标卡尺测量双侧衬垫厚度。用 Western 印迹法观察病变关节滑膜中 cPLA2α 信号的表达。用苏木精-伊红(HE)染色法观察大鼠踝关节滑膜的病理变化。用酶联免疫吸附法检测白细胞介素(IL)-1β、IL-10、前列腺素 E2(PGE2)和白三烯 B4(LTB4):结果:艾灸提高了 Annexin 1 的水平,减轻了 RA 大鼠的炎症反应。结果:艾灸增加了 Annexin 1 的表达,抑制了 cPLA2α 的磷酸化表达,降低了血清中 IL-1β、PGE2 和 LTB4 的水平,增加了 IL-10 的水平。艾灸干预Annexin 1慢病毒后的RA大鼠血清中IL-1β、PGE2、LTB4和IL-10水平几乎没有变化:结论:艾灸增强了对cPLA2α信号通路的负调控,增加了滑膜Annexin 1的表达,抑制了cPLA2α信号通路,间接抑制了下游炎症因子的表达,起到了减轻炎症的作用。
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